Engraftment of dogs with Ia-positive marrow cells isolated by avidin-
biotin immunoadsorption
RJ Berenson, WI Bensinger, D Kalamasz, F Schuening, HJ Deeg, T Graham and R Storb
Previous work has shown failure of engraftment in lethally irradiated dogs
when autologous marrow was depleted of Ia-positive cells with an anti-Ia
antibody and complement before infusion. In the current study, we have
utilized an avidin-biotin immunoadsorption procedure to obtain a population
of highly enriched Ia-positive cells for autologous bone marrow
transplantation in dogs given lethal irradiation. Dog marrow cells (2.4 to
7.0 X 10(9) cells) that contained 8.6% to 19.9% Ia- positive cells were
treated successively with monoclonal antibody 7.2, which reacts with a
framework determinant of Ia-antigen, and biotin- conjugated goat antimouse
immunoglobulin. These treated cells were passed over a column of
avidin-Biogel (polyacrylamide) and the adherent cells removed by mechanical
agitation. Seven lethally irradiated dogs were transplanted with 5.9 to
33.4 X 10(6) recovered adherent cells per kilogram of which 69.0% to 88.0%
were Ia-positive. All dogs had hematologic recovery; six are alive and well
with durable engraftment and one died on day 15 posttransplant. They are
immunologically normal as determined by lymph node and bone marrow
biopsies, lymphocyte function, and immunophenotyping of peripheral blood
and bone marrow cells. These data provide further evidence that canine
hematopoietic stem cells express Ia-like antigens and that these cells are
capable of complete hematopoietic and immunologic reconstitution in an
autologous model.
Volume 69,
Issue 5,
pp. 1363-1367,
05/01/1987
Copyright © 1987 by The American Society of Hematology
